Scanning electron microscopy as an auxiliary method in the study of exhumed bones

Scanning electronmicroscopy (SEM) has been used in forensic science in many ways. The reports of cases in which SEM has been used as an auxiliary method in the investigation of exhumed bones are rare. In this article, we report an exhumation that was made to determine if a seized weapon could have been used in a homicide. We used SEM to analyze a fracture in the interior of the skull of the victim. The findings described in this article showed us that it is possible to develop new researches in this field. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

Homocysteine and cysteine concentrations are modified by recent exposure to environmental air pollution in Sao Paulo, Brazil

Millions of people worldwide are affected by anthropogenic air pollution derived from the combustion of fossil fuels. In this work, we tested the effects of fetal, lactation and post-weaning ambient air pollution exposure on total homocysteine (tHcy) concentrations and on a downstream pathway element, the plasma cysteine (Cys) concentration. Two similar exposure chambers (polluted and filtered chamber) were located near an area with heavy traffic in Sao Paulo, Brazil, and male Swiss mice were housed there from the pre-natal period until 3 months of age. Groups during fetal, lactation and adult periods of exposure were apportioned, and tHcy and Cys plasma concentrations were assessed when the animals were 3 months old. In our study, both the tHcy and Cys concentrations were decreased in groups that spent their final stage of life in polluted chambers, suggesting recent alterations in tHcy and Cys concentrations due to air pollution exposure. The possible relationship of these data with cardiovascular dysfunction is still a matter of controversy in animals; nevertheless, epigenetic mechanisms emerge as a possible issue to consider in the investigation of the link between air pollution and Hcy measurement. (C) 2009 Elsevier Inc. All rights reserved.

Immunization with pVAXhsp65 Decreases Inflammation and Modulates Immune Response in Experimental Encephalomyelitis

Background: A DNA vaccine (pVAXhsp65) containing the gene of a heat-shock protein (hsp65) from Mycobacterium leprae showed high immunogenicity and protective efficacy against tuberculosis in BALB/c mice. A possible deleterious effect related to autoimmunity needed to be tested because hsp65 is highly homologous to the correspondent mammalian protein. In this investigation we tested the effect of a previous immunization with DNAhsp65 in the development of experimental autoimmune encephalomyelitis (EAE), a rat model of multiple sclerosis. Methods: Female Lewis rats were immunized with 3 pVAXhsp65 doses by intramuscular route. Fifteen days after the last DNA dose the animals were evaluated for specific immunity or submitted to induction of EAE. Animals were evaluated daily for weight loss and clinical score, and euthanized during the recovery phase to assess the immune response and inflammatory infiltration at the central nervous system. Results: Immunization with pVAXhsp65 induced a specific immune response characterized by production of IgG(2b) anti-hsp65 antibodies and IFN-gamma secretion. Previous immunization with pVAXhsp65 did not change EAE clinical manifestations (weight and clinical score). However, the vaccine clearly decreased brain and lumbar spinal cord inflammation. In addition, it downmodulated IFN-gamma and IL-10 production by peripheral lymphoid organs. Conclusion: Our data demonstrated that this vaccine does not trigger a deleterious effect on EAE development and also points to a potential protective effect. Copyright (C) 2010 S. Karger AG, Basel

Role of Ulnar Nerve Sonography in Leprosy Neuropathy With Electrophysiologic Correlation

Objective. The purpose of this study was to evaluate the diagnostic usefulness of ulnar nerve sonography in leprosy neuropathy with electrophysiologic correlation. Methods. Twenty-one consecutive patients with leprosy (12 men and 9 women; mean age +/- SD, 47.7 +/- 17.2 years) and 20 control participants (14 men and 6 women; mean age, 46.5 +/- 16.2 years) were evaluated with sonography. Leprosy diagnosis was established on the basis of clinical, bacteriologic, and histopathologic criteria. The reference standard for ulnar neuropathy in this study was clinical symptoms in patients with proven leprosy The sonographic cross-sectional areas (CSAs) of the ulnar nerve in 3 different regions were obtained. Statistical analyses included Student t tests and receiver operating characteristic curve analysis. Results. The CSAs of the ulnar nerve were significantly larger in the leprosy group than the control group for all regions (P < .01). Sonographic abnormalities in leprosy nerves included focal thickening (90.5%), hypoechoic areas (81%), loss of the fascicular pattern (33.3%), and focal hyperechoic areas (4.7%). Receiver operating characteristic curve analysis showed that a maximum CSA cutoff value of 9.8 mm(2) was the best discriminator (sensitivity, 0.91; specificity, 0.90). Three patients with normal electrophysiologic findings had abnormal sonographic findings. Two patients had normal sonographic findings, of which 1 had abnormal electrophysiologic findings, and the other refused electrophysiologic testing. Conclusions. Sonography and electrophysiology were complementary for identifying ulnar nerve neuropathy in patients with leprosy, with clinical symptoms as the reference standard. This reinforces the role of sonography in the investigation of leprosy ulnar neuropathy.

Deregulation of LOXL1 and HTRA1 Gene Expression in Endometriosis

Endometriosis is a gynecologic disease characterized by the presence of endometrial tissue outside the uterine cavity. Although 15% of the female population in reproductive age is affected by endometriosis, its pathogenesis remains unclear. According to the most accepted pathogenesis hypothesis, endometrial fragments from the menstrual phase are transported through the uterine tubes to the peritoneal cavity, where they undergo implantation and growth, invading adjacent tissues. However, the establishment of the disease requires that endometrial cells present molecular characteristics favoring the onset and progression of ectopic implantation. In this investigation, we analyzed the differential gene expression profiles of peritoneal and ovarian endometriotic lesions compared to the endometrial tissue of nonaffected women using rapid subtraction hybridization (RaSH). In our study, this method was applied to samples of endometriotic lesions from affected women and to biopsies of endometrium of healthy women without endometriosis, where we could identify 126 deregulated genes. To evaluate the expression of genes found by RaSH method, we measured LOXL1, HTRA1, and SPARC genes by real-time polymerase chain reaction. Significant different expression was obtained for HTRA1 and LOXL1, upregulated in the ectopic endometrium, suggesting that these genes are involved in the physiopathology of endometriosis and may favor the viability of endometrial cells at ectopic sites.

Repeatability of maximal voluntary force and of surface EMG variables during voluntary isometric contraction of quadriceps muscles in healthy subjects

The repeatability of initial values and rate of change of EMG signal mean spectral frequency (MNF), average rectified values (ARV), muscle fiber conduction velocity (CV) and maximal voluntary contraction (MVC) was investigated in the vastus medialis obliquus (VMO) and vastus lateralis (VL) muscles of both legs of nine healthy male subjects during voluntary, isometric contractions sustained for 50 s at 50% MVC. The values of MVC were recorded for both legs three times on each day and for three subsequent days, while the EMG signals have been recorded twice a day for three subsequent days. The degree of repeatability was investigated using the Fisher test based upon the ANalysis Of VAriance (ANOVA), the Standard Error of the Mean (SEM) and the Intraclass Correlation Coefficient (ICC). Data collected showed a high level of repeatability of MVC measurement (normalized SEM from 1.1% to 6.4% of the mean). MNF and ARV initial values also showed a high level of repeatability (ICC > 70% for all muscles and legs except right VMO). At 50% MVC level no relevant pattern of fatigue was observed for the VMO and VL muscles, suggesting that other portions of the quadriceps might have contributed to the generated effort. These observations seem to suggest that in the investigation of muscles belonging to a multi-muscular group at submaximal level, the more selective electrically elicited contractions should be preferred to voluntary contractions. (C) 2001 Elsevier Science Ltd. All rights reserved.

Development and reorganization of corticospinal projections in EphA4 deficient mice

The Eph family of receptor tyrosine kinases and their ligands, the ephrins, are important regulators of axon guidance and cell migration in the developing nervous system. Inactivation of the EphA4 gene results in axon guidance defects of the corticospinal tract, a major descending motor pathway that originates in the cortex and terminates at all levels of the spinal cord. In this investigation, we report that although the initial development of the corticospinal projection is normal through the cortex, internal capsule, cerebral peduncle, and medulla in the brain of EphA4 deficient animals, corticospinal axons exhibit gross abnormalities when they enter the gray matter of the spinal cord. Notably, many corticospinal axons fail to remain confined to one side of the spinal cord during development and instead, aberrantly project across the midline, terminating ipsilateral to their cells of origin. Given the possible repulsive interactions between EphA4 and one of its ligands, ephrinB3, this defect could be consistent with a loss of responsiveness by corticospinal axons to ephrinB3 that is expressed at the spinal cord midline. Furthermore, we show that EphA4 deficient animals exhibit ventral displacement of the mature corticospinal termination pattern, suggesting that developing corticospinal axons, which may also express ephrinB3, fail to be repelled from areas of high EphA4 expression in the intermediate zone of the normal spinal cord. Taken together, these results suggest that the dual expression of EphA4 on corticospinal axons and also within the surrounding gray matter is very important for the correct development and termination of the corticospinal projection within the spinal cord. J. Comp. Neurol. 436: 248-262, 2001. (C) 2001 Wiley-Liss, Inc.

Erythroid differentiation and protoporphyrin IX down-regulate frataxin expression in Friend cells: characterization of frataxin expression compared to molecules involved in iron metabolism and hemoglobinization

Friedreich ataxia (FA) Is caused by decreased frataxin expression that results in mitochondrial iron (Fe) overload. However, the role of frataxin in mammalian Fe metabolism remains unclear. In this investigation we examined the function of frataxin in Fe metabolism by implementing a well-characterized model of erythroid differentiation, namely, Friend cells induced using dimethyl sulfoxide (DMSO). We have characterized the changes in frataxin expression compared to molecules that play key roles in Fe metabolism (the transferrin receptor [TfR] and the Fe transporter Nramp2) and hemoglobinization (beta-globin). DMSO induction of hemoglobinization results in a marked decrease in frataxin gene (Frda) expression and protein levels. To a lesser extent, Nramp2 messenger RNA (mRNA) levels were also decreased on erythroid differentiation, whereas TfR and beta-globin mRNA levels increased. Intracellular Fe depletion using desferrioxamine or pyridoxal isonicotinoyl hydrazone, which chelate cytoplasmic or cytoplasmic and mitochondrial Fe pools, respectively, have no effect on frataxin expression. Furthermore, cytoplasmic or mitochondrial Fe loading of induced Friend cells with ferric ammonium citrate, or the heme synthesis inhibitor, succinylacetone, respectively, also had no effect on frataxin expression. Although frataxin has been suggested by others to be a mitochondrial ferritin, the lack of effect of intracellular Fe levels on frataxin expression is not consistent with an Fe storage role. Significantly, protoporphyrin IX down-regulates frataxin protein levels, suggesting a regulatory role of frataxin in Fe or heme metabolism. Because decreased frataxin expression leads to mitochondrial Fe loading in FA, our data suggest that reduced frataxin expression during erythroid differentiation results in mitochondrial Fe sequestration for heme biosynthesis. (C) 2002 by The American Society of Hematology.

Use of confocal microscopy to follow the development of penetrative hyphae during growth of Rhizopus oligosporus in an artificial solid-state fermentation system

Two methods were compared for determining the concentration of penetrative biomass during growth of Rhizopus oligosporus on an artificial solid substrate consisting of an inert gel and starch as the sole source of carbon and energy. The first method was based on the use of a hand microtome to make sections of approximately 0.2- to 0.4-mm thickness parallel to the substrate surface and the determination of the glucosamine content in each slice. Use of glucosamine measurements to estimate biomass concentrations was shown to be problematic due to the large variations in glucosamine content with mycelial age. The second method was a novel method based on the use of confocal scanning laser microscopy to estimate the fractional volume occupied by the biomass. Although it is not simple to translate fractional volumes into dry weights of hyphae due to the lack of experimentally determined conversion factors, measurement of the fractional volumes in themselves is useful for characterizing fungal penetration into the substrate. Growth of penetrative biomass in the artificial model substrate showed two forms of growth with an indistinct mass in the region close to the substrate surface and a few hyphae penetrating perpendicularly to the surface in regions further away from the substrate surface. The biomass profiles against depth obtained from the confocal microscopy showed two linear regions on log-linear plots, which are possibly related to different oxygen availability at different depths within the substrate. Confocal microscopy has the potential to be a powerful tool in the investigation of fungal growth mechanisms in solid-state fermentation. (C) 2003 Wiley Periodicals, Inc.

Residents’ perceptions of tourism impacts in Guimarães (Portugal): a cluster analysis

In this investigation, a cluster analysis was used to separate Guimara˜es (Portugal) residents into clusters according to their perceptions of the impacts of tourism development. This approach is uncommonly applied to Portugal data and is even rarer for world heritage sites. The world heritage designation is believed to make an area more attractive to tourists. The clustering procedure analysed 400 data observations from a Guimara˜es resident survey and revealed the existence of three clusters: the Sceptics, the Moderately Optimistic and the Enthusiasts. The results were consistent with the empirical literature’s results, with the emergent nature of the destination found to be relevant. The fact that tourism is relatively recent in this destination has its major reflex in the devaluation by most of the residents of the negative impacts of tourism development.

Psicologia das motivações ajurídicas do sentenciar : a emergência do saber em detrimento do poder

RESUMO:A pretensão deste trabalho reside na indagação sobre a eventual existência de um acervo de motivações ajurídicas que fundamentem as decisões judicias, erigindo como actor principal o Aplicador da Lei. Para tanto – recorreu-se à utilização de um instrumento construído pela autora, em 2005 – a Grelha de Observação das Motivações Ajurídicas do Sentenciar – destinado a captar aspectos considerados principais na comunicação dos protagonistas do processo de criminalização secundária, quer ao nível da comunicação verbal (C.V.) quer no concerne à comunicação não verbal (C.N.V.), verificando, depois, em presença das decisões, as razões justificativas evidenciadas. E que constituíram o suporte ajurídico de cada sentença. Assim, realizou-se um trabalho de observação, tendo a Grelha de Observação sido aplicada por licenciados em Psicologia, estudantes do 2º ciclo de Psicologia Forense e da Exclusão Social (Universidade Lusófona de Humanidades e Tecnologias), e alunos da Faculdade de Direito (Universidade Nova de Lisboa), que operaram em três tribunais criminais de Lisboa A amostra é constituída por 25 julgamentos, com um total de 64 personagens judiciais. Os resultados, ainda que meramente indicativos, demonstram uma tendência para a validade e consistência interna da grelha, indiciando também a verificação de motivações ajurídicas nas decisões judiciais. ABSTRACT: The intention of this work resides in the investigation of the eventual existence of an amount of non judicial motivations which support judicial decisions, where the law applicator arises as the main actor. For such - the use of an instrument built by the author in 2005 was resorted to - the Observation grid of the non judicial motivations of the sentencing - destined to capture aspects considered primary in the communication of the key players of the secondary criminalization process, in both the verbal communication level and in what concerns non verbal communication, later verifying, in the presence of the decisions, the evidenced justifiable reasons, which established the non judicial support of each sentence. Thus, an observational work was performed, having been applied the observation grid by Psychology graduates, students of the masters in Forensic Psychology and of Social Exclusion (Lusófona University of Humanities and Technology), as well as students of the Faculty of Law (New University of Lisbon), which operated in three criminal courts in Lisbon. The sample consists of 25 trials, with a total of 64 judicial characters. The results, even if only indicative, show a trend towards internal consistency and validity of the grid, also indicating the verification of non judicial motivations in judicial decisions.

Evaluation of 99mTc-Sestamibi as a potential tool to investigate PgP activity in inflammation

Introduction: In the XXI Century ’s Society the scientific investigation process has been growing steadily , and the field of the pharmaceutical research is one of the most enthusiastic and relevant . Here, it is very important to correlate observed functional alterations with possibly modified drug bio distribution patterns . Cancer, inflammation and inf ection are processes that induce many molecular intermediates like cytokines, chemokines and other chemical complexes that can alter the pharmacokinetics of many drugs. One cause of such changes is thought to be the modulator action of these complexes in t he P - Glyco p rotein activity, because they can act like inducers/inhibitors of MDR - 1 expression. This protein results from the expression of MDR - 1 gene, and acts as an ATP energy - dependent efflux pump, with their substrates including many drugs , like antiretrovirals, anticancers, anti - infectives, immunosuppressants, steroids or opioids . Objectives: Because of the lack of methods to provide helpful information in the investigation of in vivo molecular changes in Pgp activity during infection/infl ammation processes, and its value in the explanation of the altered drug pharmacokinetic, this paper want to evaluate the potential utility of 99m Tc - Sestamibi scintigraphy during this kind of health sciences investigation. Although the a im is indeed to create a technique to the in vivo study of Pgp activity, this preliminary Project only reaches the in vitro study phase, assumed as the first step in a n evaluation period for a new tool development. Materials and Methods: For that reason , we are performing in vitro studies of influx and efflux of 99m Tc - Sestamibi ( that is a substrate of Pgp) in hepatocytes cell line (HepG2). We are interested in clarify the cellular behavior of this radiopharmaceutical in Lipopolysaccharide(LPS) stimulated cells ( well known in vitro model of inflammation) to possibly approve this methodology. To validate the results, the Pgp expression will be finally evaluated using Western Blot technique. Results: Up to this moment , we still don’t have the final results, but we have already enough data to let us believe that LPS stimulation induce a downregulation of MDR - 1, and consequently Pgp, which could conduce to a prolonged retention of 99m Tc - Sestamibi in the inflamed cells . Conclusions: If and when this methodology demonstrate the promising results we expect, one will be able to con clude that Nuclear Medicine is an important tool to help evidence based research also on this specific field .

Prevalence of anemia and associated factors in older adults: evidence from the SABE Study

OBJECTIVE To assess the prevalence of anemia and associated factors in older adults. METHODS The prevalence and factors associated with anemia in older adults were studied on the basis of the results of the Saúde, Bem-Estar e Envelhecimento (SABE – Health, Welfare and Aging) study. A group of 1,256 individuals were interviewed during the third wave of the SABE study performed in Sao Paulo, SP, in 2010. The study included 60.4% females; the mean age of the participants was 70.4 years, and their average education was 5.3 years. The dependent variable was the presence of anemia (hemoglobin levels: 12 g/dL in women and 13 g/dL in men). Descriptive analysis and hierarchical logistic regression were performed. The independent variables were as follows: a) demographics: gender, age, and education and b) clinical characteristics: self-reported chronic diseases, presence of cognitive decline and depression symptoms, and body mass index. RESULTS The prevalence of anemia was 7.7% and was found to be higher in oldest adults. There was no difference between genders, although the hemoglobin distribution curve in women showed a displacement toward lower values in comparison with the distribution curve in men. Advanced age (OR = 1.07; 95%CI 0.57;1.64; p < 0.001), presence of diabetes (OR = 2.30; 95%CI 1.33;4.00; p = 0.003), cancer (OR = 2.72; 95%CI 1.2;6.11; p = 0.016), and presence of depression symptoms (OR = 1.75; 95%CI 1.06;2.88; p = 0.028) remained significant even after multiple analyses. CONCLUSIONS The prevalence of anemia in older adults was 7.7% and was mainly associated with advanced age and presence of chronic diseases. Thus, anemia can be an important marker in the investigation of health in older adults because it can be easily diagnosed and markedly affects the quality of life of older adults.

Risk factors and prevalence of antibodies against hepatitis A virus (HAV) in children from day-care centers, in Goiania, Brazil

A seroepidemiologic survey about hepatitis A virus (HAV) infection was carried out in a group comprising 310 children, ranging in age from 3 months to 9 years, from day-care centers, in Goiania, a middle sized city in the central region of Brazil. The biomarkers employed in the investigation of previous infection include total IgG and IgM anti-HAV antibodies, and for the detection of more recent infection, IgM anti-HAV antibodies were analyzed. The study was performed in 1991 and 1992. According to the results, 69.7% of the children presented total IgG/IgM anti-HAV antibodies, with 60% of the group in the age range of 1 to 3 years. Among 10 day-care centers analyzed, the prevalence of the biomarker IgM anti-HAV was 3.2%, with an uniform distribution of the cases in the group of children ranging in age from 1 to 4 years. Multi-variate analysis was performed to investigate the sociodemographic factors that could influence the results. It was verified that the risk for the infection increased with the length of the attendance in the day-care centers, i.e., the risk for children with attendance of one year or more was 4.7 times higher, when compared with children with one month attendance (CI 95% 2.3-9.9). According to the results, hepatitis A is an endemic infection in day-care centers in the study area. The length of attendance in the day-care settings was demonstrated to be a risk factor for the HAV infection. Such findings suggest that if hepatits A vaccination becomes available as a routine policy in our region, the target group should be children under one year. Moreover, those children should receive the vaccine before they start to attend the day-care centers.

Expression of cyclooxygenase enzymes and prostaglandin E2 receptors in inflammatory airway diseases

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina